USE OF METAL COMPLEXES CONTAINING PERFLUOROALKYL AS CONTRAST AGENTS IN MAGNETIC RESONANCE IMAGING FOR REPRESENTING INTRAVASCULAR THROMBI

UTILISATION DE COMPLEXES METALLIQUES CONTENANT DU PERFLUOROALKYLE EN TANT QU'AGENTS DE CONTRASTE EN IMAGERIE PAR RESONANCE MAGNETIQUE POUR LA REPRESENTATION DE THROMBUS INTRAVASCULAIRES

English Abstract

The invention relates to the use of metal complexes containing perfluoroalkyl
as contrast agents in MR imaging for representing intravascular thrombi. Said
complexes have a critical micelle-formation concentration < 10-3 mol/l, a
hydrodynamic micelle diameter (2 Rh) > 1 nm and a proton relaxivity in plasma
(R1) > 10 l/mmol.s.

French Abstract

L'invention concerne l'utilisation, en tant qu'agents de contraste pour l'imagerie par résonance magnétique, de complexes métalliques contenant du perfluoroalkyle et présentant une concentration critique de formation de micelles inférieure à 10?-3¿ mole/l, un diamètre micellaire hydrodynamique (2 Rh) supérieur à 1 nm et une relaxivité des protons dans le plasma (R?1¿) supérieure à 10 l/mmole.s, et ce pour la représentation de thrombus intravasculaires.

Claims

42


Claims

1. Use of perfluoroalkyl-containing metal complexes that have a critical
micelle
formation concentration < 10 -3 mol/l, a hydrodynamic micelle diameter (2 Rh)
> 1 nm and a
proton relaxivity in plasma (R1) > 10 l/mmols as contrast media in MR imaging
for visualization
of intravascular thrombi.
2. Use according to claim 1, characterized in that the metal complexes are
used as MRI
contrast media for visualization of venous thrombi.
3. Use according to claim 1 or 2, wherein the metal complexes are used as MRI
contrast
media for visualization of arterial thrombi.
4. Use according to one of claims 1 to 3, wherein the metal complexes are used
as MRI
contrast media for early determination of a thrombotic occlusive disease.
5. Use according to claim 1, wherein metal complexes whose micelle formation
concentration is < 10 -4 mol/l are used.
6. Use according to claim 1, wherein metal complexes whose hydrodynamic
micelle
diameter is >= 3 nm, preferably > 4 nm, are used.
7. Use according to claim 1, wherein metal complexes that have a proton
relaxivity in
plasma of > 13 l/mmols, preferably > 15 l/mmols, are used.
8. Use according to one of claims 1 to 7, wherein as perfluoroalkyl-containing
metal
complexes, the compounds of general formula I
R F-L-K ~I
in which
R F is a perfluorinated, straight-chain or branched carbon chain with formula -
C n F2n E,
in which
E represents a terminal fluorine, chlorine, bromine, iodine or hydrogen atom
and n stands for numbers 4-30,
L means a direct bond, a methylene group, an -NHCO group, a group


93


Image
whereby p means the numbers 0 to 10, and q and n, independently of one
another, mean numbers 0 or 1, and
R a is a hydrogen atom, a methyl group, a benzyl group, a phenyl
group, a -CH2-OH group, a CH2OCH3 group, a -CH2-CO2H group
or a C2-C15 chain, which optionally is interrupted by 1 to 3 oxygen
atoms, 1 to 2 >CO groups or an optionally substituted aryl group
and/or is substituted with 1 to 4 hydroxyl groups, 1 to 2 C1-C4
alkoxy groups, 1 to 2 carboxy groups, or a group -SO3H-,
or is a straight-chain, branched, saturated or unsaturated C2-C30 carbon
chain, which optionally contains 1 to 10 oxygen atoms, 1 to 3 -NR a
groups, 1 to 2 sulfur atoms, a piperazine, a -CONR a group, one to six -
NR a CO groups, an -SO2 group, an -NR a-CO2 group, 1 to 2 CO groups, a
group
Image or 1 to 2 optionally substituted
aryls and/or is interrupted by these groups and/or is optionally substituted
with 1 to 3 -OR a groups, 1 to 2 oxo groups, 1 to 2 -NH-COR a groups, 1 to
2 -CONHR a groups, 1 to 2 -(CH2)p-CO2H groups, 1 to 2 groups -(CH2)p-
(O)q-CH2CH2-R F,
whereby
R a, R F and p and q have the above-indicated meanings, and


44


T means a C2-C10 chain, which optionally is interrupted by 1 to 2
oxygen atoms or 1 to 2 -NHCO groups,
K stands for a complexing agent or metal complex or their salts of organic
and/or
inorganic bases or amino acids or amino acid amides, specifically for a
complexing agent or complex of general formula II
Image
in which R c, R1 and B are independent of one another, and
R c has the meaning of R a or means -(CH2)m-L-R F, whereby m is 0, 1 or 2,
and L and R F have the above-mentioned meaning,
R 1, independently of one another, mean a hydrogen atom or a metal ion
equivalent of atomic numbers 22-29, 42-46 or 58-70,
B means -OR1 or
Image
whereby R1, L, R F and R c have the above-mentioned meanings, or
K stands for a complexing agent or complex of general formula III


45


Image
in which R c and R1 have the above-mentioned meanings,
R b has the meaning of R a,
or
K stands for a complexing agent or complex of general formula IV
Image
in which R1 has the above-mentioned meaning
or
K stands for a complexing agent or complex of general formula V


46


Image
in which R1 has the above-mentioned meaning, and o and q stand for number 0 or
1, and yields the sum o + q = 1,
or
K stands for a complexing agent or complex of general formula VI
Image
in which R1 has the above-mentioned meaning
or
K stands for a complexing agent or complex of general formula VII



47


Image
in which R1 and B have the above-mentioned meanings
or
K stands for a complexing agent or complex of general formula VIII
Image
in which R c and R1 have the above-mentioned meanings, and R b has the
above-mentioned meaning of R a
or
K stands for a complexing agent or complex of general formula IX
Image


48


in which R c and R1 have the above-mentioned meanings,
or
K stands for a complexing agent or complex of general formula X
Image
in which R c and R1 have the above-mentioned meanings,
or
K stands for a complexing agent or complex of general formula XI
Image
in which R1, p and q have the above-mentioned meaning, and R b has the meaning
of R a,
or
K stands for a complexing agent or complex of general formula XII


49


Image
in which L, R F and Z1 have the above-mentioned meanings,
or
K stands for a complexing agent or complex of general formula XIII
Image
in which R1 has the above-mentioned meaning,
are used.
9. Use according to claim 8, wherein the compounds of general formula I, in
which L
stands for
.alpha.-CH2-.beta.
.alpha.-CH2CH2-.beta.
.alpha.-(CH2)s-.beta. s = 3 - 15
.alpha.-CH2-O-CH2CH2-.beta.
.alpha.-CH2-(O-CH2-CH2-)t-.beta. t = 2 - 6
.alpha.-CH2-NH-CO-.beta.
.alpha.-CH2-NH-CO-CH2-N(CH2COOH)-SO2-.beta.


50


.alpha.-CH2-NH-CO-CH2-N(C2H5)-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(C10H21)-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(C6H13)-SO2-.beta.
.alpha.-CH2-NH-CO-(CH2)10-N(C2H5)SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(-CH2-C6H5)-SO2-.beta.
.alpha. CH2-NH-CO-CH2-N(-CH2-CH2-OH)SO2-.beta.
.alpha.-CH2-NHCO-(CH2)10-S-CH2CH2-.beta.
.alpha.-CH2NHCOCH2-O-CH2CH2-.beta.
.alpha.-CH2NHCO(CH2)10-O-CH2CH2-.beta.
.alpha.-CH2-C6H4-O-CH2CH2-.beta.
.alpha.-CH2-O-CH2-C(CH2-OCH2CH2-C6F13)2-CH2-OCH2-CH2-.beta.
.alpha.-CH2-NHCOCH2CH2CON-CH2CH2NHCOCH2N(C2H5)SO2C8F17
¦
CH2-CH2NHCOCH2N(C2H5)-SO2-.beta.
.alpha.-CH2-O-CH2-CH(OC10H21)-CH2-O-CH2CH2-.beta.
.alpha.-(CH2NHCO)4-CH2O-CH2CH2-.beta.
.alpha.-(CH2NHCO)3-CH2O-CH2CH2-.beta.
.alpha.-CH2-OCH2C(CH2OH)2-CH2-O-CH2CH2-.beta.
Image
.alpha.-CH2NHCOCH2N(C6NH5)-SO2-.beta.
.alpha.-NHCO-CH2-CH2-.beta.
.alpha.-NHCO-CH2-O-CH2CH2-.beta.
.alpha.-NH-CO-.beta.
.alpha.-NH-CO-CH2-N(CH2COOH)-SO2-.beta.
.alpha.-NH-CO-CH2-N(C2H5)-SO2-.beta.
.alpha.-NH-CO-CH2-N(C10H21)-SO2-.beta.
.alpha.-NH-CO-CH2-N(C6H13)-SO2-.beta.
.alpha.-NH-CO-(CH2)10-N(C2H5)-SO2-.beta.
.alpha.-NH-CO-CH2-N(-CH2-C6H5)-SO2-.beta.
.alpha.-NH-CO-CH2-N(-CH2-CH2-OH)SO2-.beta.
.alpha.-NH-CO-CH2-.beta.


51


.alpha.-CH2-O-C6H4-O-CH2CH2-.beta.
.alpha.-CH2-C6H4-O-CH2-CH2-.beta.
.alpha.-N(C2H5)-SO2-.beta.
.alpha.-N(C6H5)-SO2-.beta.
.alpha.-N(C10H21)-SOS-.beta.
.alpha.-N(C6H13)-SO2-.beta.
.alpha. N(CH2H4OH)-SO2-.beta.
.alpha. N(CH2COOH)-SO2-.beta.
.alpha.-N(CH2C6H5)-SO2-.beta.
.alpha.-N-[CH(CH2OH)2]-SO2-.beta.
.alpha.-N-[CH(CH2OH)CH(CH2OH))-SO2-.beta.
and in which .alpha. represents the binding site to the complexing agent or
metal complex K, and .beta.
represents the binding site to the fluorine radical,
are used.
10. Use according to claim 8 or 9, wherein the compounds of formula I in which
n in
formula -C n F2n E stands for numbers 4-15 and/or E in this formula means a
fluorine atom are
used.
11. Use according to one of claims 8 to 10, wherein the following compounds
are used:
-- Gadolinium complex of 10-[1-methyl-2-oxo-3-aza-5-oxo-{4-
perfluorooctylsulfonyl-piperazin-1-yl}-pentyl]-1,4,7-tris(carboxymethyl)-
1,4,7,10-
tetraazacyclododecane,
-- Gadolinium complex of 10-[2-hydroxy-4-aza-5-oxo-7-oxa-
10,10,11,11,12,12,13,13,14,14,15,15,16,16,17,17-heptadecafluoroheptadecyl]-
1,4,7-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane,
-- Gadolinium complex of 10-[2-hydroxy-4-aza-5,9-dioxo-9-{4-perfluorooctyl)-
piperazin-1-yl}-nonyl]-1,4,7-tris(carboxymethyl)-1,4,7,10-
tetraazacyclododecane,
-- Gadolinium complex of 10-[2-hydroxy-4-aza-5-oxo-7-aza-7-(perfluorooctyl-
sulfonyl)-nonyl]-1,4,7-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane,
-- Gadolinium complex of 10-[2-hydroxy-4-oxa-1H,1H,2H,3H,3H,5H,5H,6H,6H-


52


perfluorotetradecyl]-1,4,7-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane,
-- Gadolinium complex of 10-[2-hydroxy-4-aza-5-oxo-7-oxa-
10,10,11,11,12,12,13,13,14,14,15,15,16,16,17,17,18,18,19,19-henicosafluoro-
nonadecyl]-1,4,7-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane,
-- Gadolinium complex of 10-[2-hydroxy-4-aza-5-oxo-11-aza-11-
(perfluorooctylsulfonyl)-tridecyl]-1,4,7-tris(carboxymethyl)-1,4,7,10-
tetraazacyclododecane,
-- Gadolinium complex of 10-[2-hydroxy-4-aza-5-oxo-7-aza-7-
(perfluorooctylsulfonyl)-8-phenyl-octyl]-1-4-7-tris(carboxymethyl)-1,4,7,10-
tetraaza-cyclododecane.
12. Use according to one of claims 1-7, wherein as perfluoroalkyl-containing
metal
complexes, the compounds of general formula Ia
A-R F (Ia)
in which
.cndot. A is a molecule part that contains 2 to 6 metal complexes, which are
bonded
directly or via a linker to a nitrogen atom of an annular skeleton chain,
and
.cndot. R F is a perfluorinated, straight-chain or branched carbon chain with
formula
-C n F2n E, in which
E represents a terminal fluorine, chlorine, bromine, iodine or
hydrogen atom,
and n stands for numbers 4-30,
whereby molecule part A has the following structure:


53


Image
whereby
.cndot. q1 is a number 0, 1, 2 or 3,
.cndot. K stands for a complexing agent or metal complex or their salts of
organic and/or
inorganic bases or amino acids or amino acid amides,
.cndot. X is a direct bond to the perfluoroalkyl group, a phenylene group or a
C1-C10-
alkylene chain, which optionally contains 1-15 oxygen atoms, 1-5 sulfur atoms,
1-
carbonyl groups, 1-10 (NR d) groups, 1-2 NR d SO2 groups, 1-10 CONR d groups,
1 piperidine group, 1-3 SO2 groups and 1-2 phenylene groups or optionally is
substituted by 1-3 radicals R F, in which R d stands for a hydrogen atom, a
phenyl
group, benzyl group or a C1-C15 alkyl group, which optionally contains 1-2
NHCO
groups, 1-2 CO groups, or 1-5 oxygen atoms and optionally is substituted by 1-
5
hydroxy, 1-5 methoxy, 1-3 carboxy, or 1-3 R F radicals,
.cndot. V is a direct bond or a chain of general formula IIa or IIIa:




54
Image
in which
~ Re is a hydrogen atom, a phenyl group, a benzyl group or a C,-C7-alkyl
group,
which optionally is substituted with a carboxy group, a methoxy group or a
hydroxy group,
~ W is a direct bond, a polyglycol ether group with up to 5 glycol units, or a
molecule part of general formula IVa
-CH(Rh)- (IVa)
in which Rh is a C1,-C7 carboxylic acid, a phenyl group, a benzyl group or a
-(CH2)1-5-NH-K group,
~ a represents the binding to the nitrogen atom of the skeleton chain, .beta.
represents
the binding to complexing agents or metal complex K,
~ and in which variables k and m stand for natural numbers between 0 and 10,
and 1 stands for 0 or 1
and whereby
D is a CO or SO2 group,




55
are used.
13. Use according to claim 12, wherein the compounds of general formula Ia in
which q
is the number 1 are used.
14. Use according to claim 12, wherein the compounds of general formula Ia are
used, in
which molecule part X is an alkylene chain, which contains 1-10 CH2CH2O groups
or 1-5
COCH2NH groups, a direct bond or one of the following structures
Image
whereby
.gamma. binds to D, and 8 binds to RF.
15. Use according to claim 12, wherein the compounds of general formula Ia, in
which V
is a molecule part with one of the following structures




56
Image
are used.
I 6. Use according to claim 12, wherein the compounds of general formula Ia,
in which K
represents a complex of general formula Va, VIa, VIIa or VIIIa,
Image

(Va)




57
Image
are used,
whereby
R4, independently of one another, are a hydrogen atom or a metal ion
equivalent




58
of the elements of atomic numbers 23-29, 42-46 or 58-70,
~ R5 is a hydrogen atom or a straight-chain, branched, saturated or
unsaturated C1-
C30 alkyl chain, which optionally is substituted by 1-5 hydroxy, 1-3 carboxy
or 1
phenyl groups) and/or optionally is interrupted by 1-10 oxygen atoms, I
phenylene group or 1 phenylenoxy group,
~ R6 is a hydrogen atom, a straight-chain or branched C1-C7 alkyl radical, a
phenyl
radical or a benzyl radical,
~ R7 is a hydrogen atom, a methyl group or ethyl group, which optionally is
substituted by a hydroxy group or carboxy group,
~ U3 is a straight-chain, branched, saturated or unsaturated C1-C20 alkylene
group
optionally containing 1-5 imino groups, 1-3 phenylene groups, 1-3 phenylenoxy
groups, 1-3 phenylenimino groups, 1-5 amide groups, 1-2 hydrazide groups, 1 -5
carbonyl groups, 1-5 ethylenoxy groups, 1 urea group, 1 thiourea group, 1-2
carboxyalkylimino groups, 1-2 ester groups, 1-1-0 oxygen atoms, 1-5 sulfur
atoms
and/or 1-5 nitrogen atoms, and/or optionally substituted by 1-5 hydroxy
groups, 1-
2 mercapto groups, 1-5 oxo groups, 1-5 thioxo groups, 1-3 carboxy groups, 1-5
carboxyalkyl groups, 1-5 ester groups and/or 1-3 amino groups, whereby the
optionally contained phenylene groups can be substituted by 1-2 carboxy
groups,
1-2 sulfone groups or 1-2 hydroxy groups
~ T1 stands for a -CO-.beta. or -NHCO-.beta. or -NHCS-.beta. group, whereby
.beta. represents the
binding site to V.
17. Use according to claim 16, wherein the C1-C20-alkylene chain that stands
for U3
contains the groups -CH2NHCO-, -NHCOCH2O-, -NHCOCH2OC6H4-,-N(CH2COZ2H)-,
-CH2OCH2-, -NHCOCH2C6H4-, -NHCSNHC6H4-, -CH2OC6H4-, -CH2CH2O- and/or is
substituted by the groups -COOH and -CH2COOH.
18. Use according to claim 16, wherein U3 stands for a -CH2-, -CH2CH2-, -
CH2CH2CH2-,
-C6H4-, -C6H10-, -CH2C6H4-, -CH2NHCOCH2CH(CH2CO2H)-C6H4-, -CH2NHCOCH2OCH2-, or




59
-CH2NHCOCH2C6H4- group.
19. Use according to claim 12, wherein the compounds of general formula Ia in
which K
has one of the following structures:
Image
are used.
20. Use according to one of claims 12 to 19, wherein the compounds of general
formula
Ia in which the perfluoroalkyl chain RF is -C6F13, -CgF17, -C10F21, or -C12F25
are used.
21. Use according to one of claims 12 to 20, wherein the gadolinium complex of
1,4,7-
tris{ 1,4,7-tris(N-(carboxylatomethyl)-10-[N-1-methyl-3,6-diaza-2,5,8-
trioxooctane-1,8-diyl)]-




60
1,4,7,10-tetraazacyclododecane, Gd complex}-10-[N-2H,2H,4H,4H,5H,5H-3-oxa-
perfluorotridecanoyl]-1,4,7,10-tetraazacyclododecane is used.
22. Use according to one of claims 1 to 7, wherein as perfluoroalkyl-
containing metal
complexes, the compounds of general formula Ib
Image
in which
K means a complexing agent or a metal complex of general formula IIb
Image
whereby
R1 stands for a hydrogen atom or a metal ion equivalent of atomic numbers
23-29, 42-46 or 58-70,
R2 and R3 stand for a hydrogen atom, a C1-C7-alkyl group, a benzyl group,
a phenyl group, -CH2OH or -CH2-OCH3,
U2 stands for radical L1, whereby L1 and U2, independently of one
another, can be the same or different, however,
A1 means a hydrogen atom, a straight-chain or branched C1-C3o alkyl group,
which
optionally is interrupted by 1-15 oxygen atoms, and/or optionally is
substituted




61

with 1-10 hydroxy groups, 1-2 COOH groups, a phenyl group, a benzyl group
and/or 1-5 -OR9 groups, with R9 in the meaning of a hydrogen atom or a C1-C7
alkyl radical, or -L1-RF,
L1 means a straight-chain or branched C1-C30-alkylene group, which optionally
is
interrupted by 1-10 oxygen atoms, 1-5 -NH-CO groups, 1-5 -CO-NH groups, by a
phenylene group optionally substituted by a COOH-group, 1-3 sulfur atoms, 1-2
-N(B1)-SO2 groups, and/or 1-2 -SO2-N(B1)- groups with B1 in the meaning of A1,
and/or optionally is substituted with radical RF, and
RF means a straight-chain or branched perfluorinated alkyl radical of formula
CnF20E,
whereby n stands for numbers 4-30, and
E stands for a terminal fluorine atom, chlorine atom, bromine atom,
iodine atom or a hydrogen atom,
and optionally present acid groups optionally can be present as salts of
organic and/or inorganic
bases or amino acids or amino acid amides,
are used.
23. Use according to claim 22, wherein the compounds of general formula Ib, in
which
R2, R3 and R9, independently of one another, mean hydrogen or a C1-C4 alkyl
group, are used.
24. Use according to claim 22, wherein the compounds of general formula Ib, in
which
A' means hydrogen, a C1-C15 alkyl radical,
the radicals
C2H4-O-CH3,C3H6-O-CH8,
C3H4-O-(C2H4-O)-C2H4-OH,
C2H4-O-(C2H4-O)-C2H4-(OCH3),
C2H4OH, C3H6OH, C4H8OH, C5H10OH, C6H12OH, C7H14OH,
CH(OH)CH2OH,
CH(OH)CH(OH)CH2OH, CH2[CH(OH)]u1,CH2OH,
CH[CH2(OH)]CH(OH)CH2OH,
C2H4CH(OH)CH2OH,
(CH2)5 COOH,


62
C2H4-O-(C2H4-O)t-CH2COOH or
C2H4-O-(C2H4-O)t-C2H4-C n F2nE,
whereby
s stands for integers 1 to 15,
t stands for integers 0 to 13,
u1 stands for integers 1 to 10,
n stands for integers 4 to 20, and
E stands for a hydrogen, fluorine, chlorine, bromine or iodine atom, and if
possible,
the branched isomers thereof,
are used.
25. Use according to claim 22, wherein the compounds of general formula Ib, in
which
A1 means hydrogen, C1-C10 alkyl,
C2H4-O-CH3, C3H6-O-CH3,
C2H4-O-(C2H4-O)x-C2H4-OH, C2H4-O-(C2H4-O)x-C2H4-OCH3,
C2H4OH, C3H6OH,
CH2[CH(OH))y CH2OH,
CH[CH2(OH)]CH(OH)CH2OH,
(CH2)w COOH,
C2H4-O-(C2H4-O)x-CH2COOH,
C2H4-O-(C2H4-O)x-C2H4-C n F2nE,
whereby
x stands for integers 0 to 5,
y stands for integers 1 to 6,
w stands for integers 1 to 10,
n stands for integers 4 to 15, and
E stands for a fluorine atom, and, if possible, the branched isomers thereof
are used.


63

26. Use according to claim 22, wherein the compounds of general formula Ib, in
which
L1 means
.alpha.-(CH2)s-.beta.
.alpha.-CH2-CH2-(O-CH2-CH2-).UPSILON.-.beta.
.alpha.-CH2-(O-CH2-CH2-).UPSILON.-.beta.,
.alpha.-CH2-NH-CO-.beta.
.alpha.-CH2-CH2-NH-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(CH2COOH)-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(C2H5)-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(C10H21)-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(C6H13)-SO2-.beta.
.alpha.-CH2-NH-CO-(CH2)10-N(C2H5)-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(-CH2-C6H5)-SO2-.beta.
.alpha.-CH2-NH-CO-CH2-N(-CH2-CH2-OH)SO2-.beta.
.alpha.-CH2-NHCO-(CH2)10-S-CH2CH2-.beta.
.alpha.-CH2NHCOCH2-O-CH2CH2-.beta.
.alpha.-CH2-CH2NHCOCH2-O-CH2CH2-.beta.
.alpha.-CH2-(CH2-CH2-O)1-(CH2)3NHCO-CH2-O-CH2CH2-.beta.
.alpha.-CH2NHCO(CH2)10-O-CH2CH2-.beta.
.alpha.-CH2CH2NHCO(CH2)10-O-CH2CH2-.beta.
.alpha.-CH2-C6H4-O-CH2CH2-.beta.,
whereby the phenylene group 1,4 or 1,3 is linked
.alpha.-CH2-O-CH2-C(CH2-OCH2CH2-C6F13)2-CH2-OCH2-CH2-.beta.
.alpha.-CH2-NHCOCH2CH3CON-CH2CH2NHCOCH2N(C2H5)SO2C8F17.beta.
.alpha.-CH2-CH2NHCOCH2N(C2H5)-SO2-.beta.
.alpha.-CH2-O-CH2-CH(OC10H21)-CH2-O-CH2CH2-.beta.
.alpha.-(CH2NHCO)4-CH2O-CH2CH2-.beta.
.alpha.-(CH2NHCO)3-CH2O-CH2CH2-.beta.
.alpha.-CH2-OCH2C(CH2OH)2-CH2-O-CH2CH2-.beta.


64

Image

.alpha.-CH2NHCOCH2N(C6H5)-SO2-.beta.
.alpha.-NHCO-CH2-CH2-.beta.
.alpha.-NHCO-CH2-O-CH2CH2-.beta.
.alpha.-NH-CO-.beta.
.alpha.-NH-CO-CH2-N(CH2COOH)-SO2-.beta.
.alpha.-NH-CO-CH2-N(C2H5)-SO2-.beta.
.alpha.-NH-CO-CH2-N(C10H21)-SO2-.beta.
.alpha.-NH-CO-CH2-N(C6H13)-SO2-.beta.
.alpha.-NH-CO-(CH2)10-N(C2H5)-SO2-.beta.
.alpha.-NH-CO-CH2-N(-CH2-C6H5)-SO2-.beta.
.alpha.-NH-CO-CH2-N(-CH2-CH2-OH)-SO2-.beta.
.alpha.-NH-CO-CH2-.beta.
.alpha.-CH2-O-C6H4-O-CH2-CH2-.beta.
.alpha.-CH2-C6H4-O-CH2-CH2-.beta.
.alpha.-N(C2H5)-SO2-.beta.
.alpha.-N(C6H5)-SO2-.beta.
.alpha.-N(C10H21)-SO2-.beta.
.alpha.-N(C6H13)-SO2-.beta.
.alpha.-N(C2H4OH)-SO2-.beta.
.alpha.-N(CH2COOH)-SO2-.beta.
.alpha.-N(CH2C6H5)-SO2-.beta.
.alpha.-N-[CH(CH2OH)2]-SO2-.beta.
.alpha.-N-[CH(CH2OH)CH(OH)(CH2OH)]-SO2-.beta.
whereby
s stands for integers 1 to 15 and
y stands for integers 1 to 6,
are used.

27. Use according to claim 22, wherein the compounds of general formula Ib, in
which L1
means


65

.alpha.-CH2-O-CH2-.beta.
.alpha.-CH2-CH2-(O-CH2-CH2-)y-.beta.,
.alpha.-CH2-(O-CH2-CH2-)y-.beta.,
.alpha.-CH2-CH2-NH-SO2-.beta.,
.alpha.-CH2NHCOCH2-O-CH2CH2-.beta.,
.alpha.-CH2-CH2NHCOCH2-O-CH2CH2-.beta.,
.alpha.-CH2(CH2-CH2-O)y-(CH2)3NHCO-CH2-O-CH2CH2-.beta.,
.alpha.-CH2NHCO(CH2)10-O-CH2CH2-.beta.,
.alpha.-CH2CH2NHCO(CH2)10-O-CH2CH2-.beta.,
.alpha.-CH2-O-CH2-CH(OC10H21)-CH2-O-CH2CH2-.beta.,
.alpha.-CH2-O-C6H4-O-CH2-CH2-.beta.
or
.alpha.-CH2-C6H4-O-CH2-CH2-.beta.
whereby
y stands for integers 1 to 6,
are used.

28. Use according to claim 22, wherein the compounds of general formula Ib, in
which
R F means a straight-chain or branched perfluorinated alkyl radical of formula
C n F2n E, whereby n
stands for numbers 4 to 15 and E stands for a terminal fluorine atom, are
used.

29. Use according to one of claims 22 to 28, wherein the following compounds
are used:
-- 1,4,7-Tris(carboxylatomethyl)-10-(3-aza-4-oxo-hexan-5-ylic)-acid-(2,3-
dihydroxypropyl)-N-(1H,1H,2H,2H,4H,4H,5H,5H-3-oxa)-perfluorotridecyl)-
amide]-1,4,7,10-tetraazacyclododecane, gadolinium complex
-- 1,4,7-Tris(carboxylatomethyl)-10-{(3-aza-4-oxo-hexan-5-ylic)acid-N-
(3,6,9,12,15-pentaoxa)-hexadecyl)-(1H,1H,2H,2H,4H,4H,5H,5H-3-oxa)-
perfluorotridecyl]-amide}-1,4,7,10-tetraazacyclododecane, gadolinium complex
-- 1,4,7-Tris(carboxylatomethyl)-10-{(3-aza-4-oxo-hexan-5-ylic)-acid-N-5-
hydroxy-
3-oxa-pentyl)-N-(1H,1H,2H,2H,4H,4H,5H,5H-3-oxa)-perfluorotridecyl]-amide}-
1,4,7,10-tetraazacyclododecane, gadolinium complex
-- 1,4,7-Tris(carboxylatomethyl)-10-{(3-aza-4-oxo-hexan-5-ylic)-acid-[N-
3,6,9,15-




66
tetraoxa-12-aza-15-oxo-C17-C26-hepta-decafluoro)hexacosyl]-amide}-1,4,7, 10-
tetraazacyclododecane, gadolinium complex
-- 1,4,7-Tris(carboxylatomethyl)-10-[(3-aza-4-oxo-hexan-5-ylic]-acid-N-(2-
methoxyethyl)-N-( 1 H, 1H,2H,2H,4H,4H,5H,5H-3-oxa)-perfluorotridecyl]-amide}-
1,4,7,10-tetraazacyclododecane, gadolinium complex.
30. Use according to one of claims 1 to 7, wherein as perfluoroalkyl-
containing metal
complexes, the compounds with sugar radicals of general formula Ic
Image
in which
R represents a mono- or oligosaccharide radical bonded by the 1-OH- or 1-SH-
position,
RF is a perfluorinated, straight-chain or branched carbon chain with the
formula
-CnF2nE, in which E represents a terminal fluorine, chlorine, bromine, iodine
or
hydrogen atom, and n stands for numbers 4-30,
K stands for a metal complex of general formula IIc,
Image
in which
R1 means a hydrogen atom or a metal ion equivalent of atomic numbers 23-29, 42-
46




67
or 58-70,
provided that at least two R1 stand for metal ion equivalents,
R2 and R3, independently of one another, represent hydrogen, C1-C7-alkyl,
benzyl, phenyl,
-CH2OH or -CH2OCH3, and
U represents -C6H4-O-CH2-.omega., -(CH2)1-5-.omega., a phenylene group, -CH2-
NHCO-CH2-
CH(CH2COOH)-C6H4-.omega., -C6H4-(OCH2CH2)0-1-N(CH2COOH)-CH2-.omega., or a C1-
C12-alkylene group or C7-C12-C6H4-O group optionally interrupted by one or
more
oxygen atoms, 1 to 3 -NHCO groups or 1 to 3 -CONH groups and/or substituted
with 1 to 3 -(CH2)0-5 COOH groups, whereby .omega. stands for the binding site
to
-CO-,
or
of general formula IIIc
Image
in which R1 has the above-mentioned meaning, R4 represents hydrogen or a metal
ion equivalent
mentioned under R1, and U1 represents -C6H4-O-CH2-.omega., whereby .omega.
means the binding
site to -CO-,
or of general formula IVc




68
Image
in which R1 and R2 have the above-mentioned meaning
or of general formula VcA or VcB
Image
in which R1 has the above-mentioned meaning,
or of general formula VIc
Image
in which R1 has the above-mentioned meaning,




69
or of general formula VIII
Image
in which R1 has the above-mentioned meaning, and
U1 represents -C6H4-O-CH2-.omega., whereby .omega. means the binding site to -
CO-
or of general formula VIIIc
Image
in which R1 has the above-mentioned meaning,
and in radical K, optionally present free acid groups optionally can be
present as salts of
organic and/or inorganic bases or amino acids or amino acid amides,
G for the case that K means metal complexes IIc to VIII represents a radical
that is
functionalized in at least three places and is selected from the following
radicals
a) to j)




70

Image




71

Image





72

Image




73

Image
and
G ~for the case that K means metal complex VIIIc represents a radical that is
functionalized in at least three places and is selected from k) or l),
Image




74

Image
whereby .alpha. means the binding site of G to complex K, .beta. is the
binding site of G to radical
Y, and .gamma. represents the binding site of G to radical Z,
Y ~means -CH2, .delta.-(CH2)(1-5)CO-.beta., .beta.-(CH2)(1-5)CO-.delta.,
.delta.-CH2-CHOH-CO-.beta. or .delta.-
CH(CHOH-CH2OH)-CHOH-CHOH-CO-.beta., whereby .delta. represents the binding
site
to sugar radical R, and .beta. is the binding site to radical G,
Z ~stands for
Image
.gamma.-COCH2-N(C2H5)-SO2-.epsilon.,
.gamma.-COH2-O-(CH2)2-SO2-.epsilon.,
Image~~~
or
.gamma. - NHCH2CH2-O-CH2CH2-..xi.,~~
whereby .gamma. represents the binding site of Z to radical G, and .xi., means
the binding
site of Z to perfluorinated radical R F
and
l 1, m 1, independently of one another, mean integer 1 or 2, and


75

p1 means integers 1 to 4,
are used.

31. Use according to claim 30, wherein the compounds of general formula Ic, in
which R
represents a monosaccharide radical with 5 to 6 C atoms or its deoxy compound,
preferably
glucose, mannose or galactose, are used.

32. Use according to claim 30, wherein the compounds of general formula Ic, in
which
R2 and R3, independently of one another, mean hydrogen or C1-C4 alkyl and/or E
in formula
-C n F2n E means a fluorine atom, are used.

33. Use according to claim 30, wherein the compounds of general formula Ic, in
which G
represents lysine radical (a) or (b), are used.

34. Use according to claim 30, wherein the compounds of general formula Ic are
used, in
which Z means
Image
whereby .gamma. represents the binding site of Z to radical G, and .xi., means
the binding site of Z to
perfluorinated radical R F, and/or Y means .delta.-CH2CO-.beta., whereby
.delta. represents the binding site to
sugar radical R, and .beta. represents the binding site to radical G.

35. Use according to claim 30, wherein the compounds of general formula Ic, in
which U
in metal complex K represents -CH2- or -C6H4-O-CH2-.omega., whereby .omega.
stands for the binding site
to -CO-, are used.

36. Use according to claim 30, wherein the gadolinium complex of 6-N-[ 1,4,7-
tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-10-N-(pentanoyl-3-aza-4-
oxo-5-methyl-
5-yl)]-2-N-[1-O-.alpha.-D-carbonylmethyl-mannopyranose]-L-lysine-[1-(4-
perfluorooctylsulfonyl)-
piperazine]-amide is used.

37. Use according to one of claims 1 to 7, wherein as perfluoroalkyl-
containing metal
complexes, the compounds with polar radicals of general formula Id


76

Image
in which
R F is a perfluorinated, straight-chain or branched carbon chain with formula -
C n F2n E,
in which E represents a terminal fluorine, chlorine, bromine, iodine or
hydrogen
atom, and n stands for numbers 4-30,
K ~stands for a metal complex of general formula IId,
Image
in which
R1 ~means a hydrogen atom or a metal ion equivalent of atomic numbers 23-29,
42-46
or 58-70,
provided that at least two R1 stand for metal ion equivalents,
R2 and R3, independently of one another, represent hydrogen, C1-C7 alkyl,
benzyl, phenyl,
-CH2OH or -CH2OCH3, and
U ~represents -C6H4-O-CH2-.omega.-, -(CH2)1-5-.omega., a phenylene group, -CH2-
NHCO-CH2-
CH(CH2COOH)-C6H4-.omega.-, -C6H4-(OCH2CH2)0-1-N(CH2COOH)-CH2-.omega., or a C1-
C12 alkylene group or C7-C12-C6H4-O group optionally interrupted by one or
more
oxygen atoms, 1 to 3 -NHCO groups, 1 to 3 -CONH groups and/or substituted
with 1 to 3 -(CH2)0-5COOH groups, whereby .omega. stands for the binding site
to -CO-,



77

or
of general formula IIId
Image
in which R1 has the above-mentioned meaning, R4 represents hydrogen or a metal
ion equivalent
mentioned under R1, and U1 represents -C6H4-O-CH2-.omega.-, whereby .omega.
means the binding site to
-CO-,
or
of general formula IVd
Image
in which R1 and R2 have the above-mentioned meaning,
or of general formula VdA or VdB




78
Image

in which R1 has the above-mentioned meaning,
or of general formula VId
Image
in which R1 has the above-mentioned meaning,
or of general formula VIId
Image
in which R1 has the above-mentioned meaning, and
U1 represents -C6H4-O-CH2-.omega.-, whereby .omega. means the binding site to -
CO-,
and in radical K, optionally present free acid groups optionally can be
present as salts of
organic and/or inorganic bases or amino acids or amino acid amides,




79

G represents a radical that is functionalized in at least three places and is
selected
from the following radicals a) to i)
Image



80

Image



81

Image
whereby .alpha. means the binding site of G to complex K, .beta. is the
binding site of G to radical R, and
.gamma. represents the binding site of G to radical Z
Z stands for
Image




82

.gamma.-C(O)CH2O(CH2)2-.epsilon.,
whereby .gamma. represents the binding site of Z to radical G, and .xi. means
the binding
site of Z to perfluorinated radical R f,
R~represents a polar radical that is selected from complexes K of general
formulas
IId to VIId, whereby R1 here means a hydrogen atom or a metal ion equivalent
of
atomic numbers 20, 23-29, 42-46 or 58-70,
and radicals R2, R3, R4, U and U1 have the above-indicated meaning,
or
R ~means the folic acid radical
or
R ~means a carbon chain with 2-30 C atoms that is bonded to radical G via -CO-
or
SO2- or a direct bond to radical G, and is straight or branched, saturated or
unsaturated, optionally interrupted by 1-10 oxygen atoms, 1-5 -NHCO groups, 1-
5
-CONH groups, 1-2 sulfur atoms, 1-5 -NH groups or 1-2 phenylene groups, which
optionally can be substituted with 1-2 OH groups, 1-2 NH2 groups, 1-2
-COOH groups, or 1-2 -SO3H groups,
or
optionally substituted with 1-8 OH groups, 1-5 -COOH groups, 1-2 SO3H groups,
1-5 NH2 groups, or 1-5 C1-C4 alkoxy groups, and
l1, m1, p2, independently of one another, mean integer 1 or 2,
are used.

38. Use according to claim 37, wherein the compounds of general formula Id, in
which K
stands for a metal complex of general formula IId, IIId, VdB or VIId, are
used.

39. Use according to claim 37, wherein the compounds of general formula Id, in
which
polar radical R has the meaning of complex K, preferably complex K of general
formulas IId,
IIId, VdA or VIId, are used.




83

40. Use according to claim 37, wherein the compounds of general formula Id, in
which
polar radical R has the following meanings:
-C(O)CH2CH2SO3H
-C(O)CH2OCH2CH2OCH2CH2OH
-C(O)CH2OCH2CH2OH
-C(O)CH2OCH2CH(OH)CH2OH
-C(4)CH2NH-C(O)CH2COOH
-C(O)CH2CH(OH)CH2OH
-C(O)CH2OCH2COOH
-SO2CH2CH2COOH
-C(O)-C6H3-(m-COOH)2
-C(O)CH2O(CH2)2-C6H3-(m-COOH)2
-C(O)CH2O-C6H4-m-SO3H
-C(O)CH2NHC(O)CH2NHC(O)CH2C3CH2COOH
-C(O)CH2OCH2CH2OCH2COOH
-C(O)CH2OCH2CH(OH)CH2O-CH2CH2OH
-C(O)CH2OCH2CH(OH)CH2OCH2-CH(OH)-CH2OH
-C(O)CH2SO3H
-C(O)CH2CH2COOH
-C(O)CH(OH)CH(OH)CH2OH
-C(O)CH2O[(CH2)2O]1-9-CH3
-C(O)CH2O[(CH2)2O]1-9)-H
-C(O)CH2OCH(CH2OH)2
-C(O)CH2OCH(CH2OCH2COOH)2
-C(O)-C6H3-(m-OCH2COOH)2
-CO-CH2O-(CH2)2O(CH2)2O-(CH2)2O(CH2)2OCH3
preferably -C(O)CH2O[(CH2)2O]4-CH3
are used.

41. Use according to claim 37, wherein the compounds of general formula Id, in
which
polar radical R is the folic acid radical, are used.

42. Use according to claim 37, wherein the compounds of general formula Id, in
which G





84~

represents lysine radical (a) or (b), are used.

43. Use according to claim 37, wherein the compounds of general formula Id, in
which U
represents group -CH2- or -C6H4-O-CH2-.omega. in metal complex K, whereby w
stands for the binding
site to -CO-, are used.

44. Use according to one of claims 37-43, wherein the gadolinium complex of
2,6-N,N'-
bis[1,4,7-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-10-(pentanoyl-
3-aza-4-oxo-5-
methyl-5-yl)]-lysine-[1-(4-perfluorooctylsulfonyl-piperazine]-amide is used.

45. Use according to one of claims 1-7, wherein as perfluoroalkyl-containing
metal
complexes, galenical formulations that contain paramagnetic, perfluoroalkyl-
containing metal
complexes of general formulas I, Ia, Ib, Ic and/or Id and diamagnetic
perfluoroalkyl-containing
substances, preferably dissolved in an aqueous solvent, are used.

46. Use according to claim 45, wherein as diamagnetic perfluoroalkyl-
containing
substances, those of general formula XX
R F-L2-B2~~ (XX)
in which R F represents a straight-chain or branched perfluoroalkyl radical
with 4 to 30 carbon
atoms, L2 stands for a linker, and B2 stands for a hydrophilic group, are
used.

47. Use according to claim 46, wherein linker L2 is a direct bond, an -SO2
group, or a
straight-chain or branched carbon chain with up to 20 carbon atoms, which can
be substituted
with one or more -OH, -COO-, or -SO3 groups and/or optionally contains one or
more -O-, -S-,
-CO-, -CONH-, -NHCO-, -CONR9-, -NR9CO-, -SO2-, -PO4- -, -NH- or -NR9 groups,
an aryl ring
or a piperazine, whereby R9 stands for a C1-to C20-alkyl radical, which in
turn can contain one or
more O atoms, and/or can be substituted with -COO- or SO3 groups.

48. Use according to claim 46, wherein hydrophilic group B2 is a mono- or
disaccharide,
one or more adjacent -COO- or -SO3 groups, a dicarboxylic acid, an isophthalic
acid, a picolinic
acid, a benzenesulfonic acid, a tetrahydropyrandicarboxylic acid, a 2,6-
pyridinedicarboxylic acid,
a quaternary ammonium ion, an aminopolycarboxylic acid, an aminodipolyethylene
glycolsulfonic acid, an aminopolyethylene glycol group, an SO2-(CH2)2-OH
group, a




85

polyhydroxyalkyl chain with at least two hydroxyl groups or one or more
polyethylene glycol
chains with at least two glycol units, whereby the polyethylene glycol chains
are terminated by an
-OH or -OCH3 group.

49. Use according to claim 45, wherein as diamagnetic perfluoroalkyl-
containing
substances, conjugates that consist of .alpha.-, .beta.- or .gamma.-
cyclodextrin and compounds of general formula
XXII:
A1-L3-R F ~~ (XXII)
in which A2 stands for an adamantane, biphenyl or anthracene molecule, L3
stands for a linker,
and R F stands for a straight-chain or branched perfluoroalkyl radical with 4
to 30 carbon atoms,
and whereby linker L3 is a straight-chain hydrocarbon chain with 1 to 20
carbon atoms, which
can be interrupted by one or more oxygen atoms, one or more CO-, SO2-, CONH-,
NHCO-,
CONR10-, NR10CO-, NH- or NR10 groups or a piperazine, whereby R10 is a C1-C5
alkyl radical,
are used.

50. Use according to claim 45, wherein as diamagnetic perfluoroalkyl-
containing
substances, those of general formula XXI:~~
R F-X1 ~~~(XXI)
in which R F represents a straight-chain or branched perfluoroalkyl radical
with 4 to 30 carbon
atoms, and X1 is a radical that is selected from the group of the following
radicals (n in this case
is a number between 1 and 10),
are used:




86

Image




87

Image




88

Image




89

Image

Description

CA 02491933 2005-O1-05
Use of PerJluoroalkyl Containing Metal Complexes as Contrast
Media in MR-Imaging for Visualization of Intravascular Thrombi
The invention relates to the subject that is characterized in the claims,
i.e., the use of
perfluoroalkyl-containing metal complexes that have a critical micelle
formation concentration
< 10-3 moll, a hydrodynamic micelle diameter (2 Rh) > 1 nm, and a proton
relaxivity in plasma
(R~) > 101/mmol~s, as contrast media in MR-imaging for visualization of
intravascular thrombi.
Thrombosis is defined as the formation of a blood clot (thrombus) in a blood
vessel and
the thus-induced constriction or clogging of this vessel. Most often,
thromboses are found in the
veins (phlebothrombosis). Here, preferably the veins of the lower half of the
body (deep femoral
and pelvic veins) are affected. Other portions of the circulatory system can
also be affected,
however: heart valves, apexei of the heart, coronary vessels, cerebral
vessels, arteries in the area
of the intestines, femoral arteries as well as veins of the leg and the
pelvis, the rectum
(hermorrhoids) and the arni. By moving the thrombus, a pulmonary embolism can
result, which
in the worst case ends in death.
Thromboses of the deep primary veins represent a significant social-medical
problem. In
Germany, 60,000 humans per year are treated for thromboses and their
consecutive symptoms.
In the USA, an acute thrombosis of the deep femoral veins of the leg and the
pelvis occurs yearly
in 48 out of a population of 100,000. About 12% of all stationary patients
develop clinically
detected deep femoral or pelvic vein thromboses. About 20 to 30% of all
general-surgery
patients and more than SO% of all patients after orthopedic/emergency surgical
intervention
suffer deep femoral vein thromboses, whereby in about 1 % of these patients, a
lung embolism
with clinical symptoms occurs (Leitlinien zu Diagnostik and Therapie in der
Gefa[3chirurgie
[Guidelines in Diagnosis and Therapy in Vascular Surgery], published by the
Board of Directors
of Dt. Ges. f. Gefa(3chirurgie; Deutscher Arzteverlag [German Society for
Vascular Surgery; '
German Physicians' Publishing House], Cologne 1998).



CA 02491933 2005-O1-05
2
The decisive mechanisms that cause a thrombosis were already described in 1856
by
Rudolf Virchow and named by him as Virchow'sTriad. In this case, this is
damage to the
vascular wall, slowing of the blood flow and an elevated blood-clotting
tendency because of a
change in the blood composition. While for the venous thrombosis
(phlebothrombosis), the
slowing of the blood flow and an elevated clotting tendency are emphasized,
damage to the
vascular wall, in most cases as a result of arteriosclerosis, with the deposit
of blood platelets
(thrombocytes) is of decisive importance in the development of the rare
arterial thrombosis.
The thrombus maintains its original form only for a few days. After a
structural
transformation, it can be transformed as a scar in its final state, and the
vessel is partially
passable again (rechanneled). The purpose of the therapy is primarily the
restoration of the blood
flow. This therapy depends on the age of the thrombus and is successful only
within the first 10
days after the thrombus develops. The restoration of the blood flow can be
carned out, on the
one hand, by a medicinal dissolution of the thrombus (thrombolysis). On the
other hand, surgical
methods are available: either the removal of the occlusion by removal of the
clot
(thrombectomy) or the bridging of the occluded vascular segment by an
angioplasty (bypass). In
the second place, the object of the therapy of the thrombosis is to prevent
additional growth of
the thrombus and to avoid late sequelae or complications.
The diagnosis of thromboses in clinical practice is mainly carned out by
imaging
processes. A very suitable method for detecting a thrombosis as well as for
determining its extent
is the radiological contrast medium study (phlebography). Drawbacks are the
exposure to
ionizing rays and the side effects that are associated with iodine-containing
contrast media. The
initial examination method in the suspicion of low femoral vein thrombosis in
many clinical
devices is therefore the color-coded duplex sonography (B-scan plus PW
Doppler), which is
extremely dependent on the examiner, however. Other non-invasive imaging
processes for
visualizing luminal vascular changes are arteriography, CT angiography and MR
angiography, as
well as methods of nuclear medicine.
Thrombi thus can be visualized by blood corpuscles that are labeled with
indium-111 as



CA 02491933 2005-O1-05
3
imaging agents (Thakur et al., Thromb. Res. 9: 345, 1976; Powers et al.,
Neurology 32: 938,
1982). The iodine isotopes J-125 and J-131 are also suitable for imaging
purposes (Pang, US
5,011,686, 1991). The technetium isotope Tc-99m is widely used as a label.
Peptides and
especially monoclonal antibodies are labeled with it (Bergen US 5,024,829,
1991; Dean et al.,
US 4,980,148, 1990: US 5,508,020, 1996; US 5,645,815, 1997; WO 00/61195; US
6,171,578,
2001; EP 1171166, 2002). Compounds that are suitable both for scintigraphy and
for MR
imaging are described by Abelman (US 5,656,600, 1997). In WO 01 /77102, DuPont
Pharmaceuticals describes conjugates from metal complexes and pyridinones,
which are suitable
as contrast media for diagnosis of thromboses with the aid of scintigraphy,
computer tomography
or MR imaging.
The literature for MR angiography for visualizing intravascular thrombi has a
broad
scope. Application WO 95/09013 describes cytogene polypeptides as complexing
agents for
paramagnetic metal ions.
In Application WO 95/24225, Nycomed named polymer complexing agents for
thrombus
imaging. Complexing agents such as DOTA or D03A are bonded to a backbone - for
example,
polylysine.
In WO 95/20603, Sandoz describes paramagnetic DTPA conjugates, which are
suitable
for the thrombus imaging.
In the Barne-Jewish Hospital Patent US 5,780,010, specifically binding (biotin-
avidin
complex) conjugates are described as contrast media for thrombus imaging.
Also, in WO
98/16256, the Burnham Institute describes radicals that specifically bind (to
integrin) and that
make possible a thrombus imaging. In these conjugates, paramagnetic complexes
of DTPA,
EDTA or DOTA are contained as signaling radicals.
Conjugates that consist of a guanidine derivative and paramagnetic complexes
are
described by 3-Dimensional Pharmaceuticals as contrast media for the thrombus
imaging in WO
01 /04117.
Conjugates that consist of complexes of DTPA, DOTA or D03A and polypeptides
are



CA 02491933 2005-O1-05
4
described by EPIX in WO 01/09188 and EP 1203026 as imaging agents for the
thrombus
imaging.
In EP 885545, Pilgrimm names superparamagnetic iron oxides as contrast media
for
thrombosis diagnosis with the aid of MRI.
The MR contrast medium that is described in WO 02/22011 for the diagnosis of
thrombi
is also particulate (USPIO).
It is disadvantageous in the case of the conjugates that in addition to the
diagnostically
active portion, they contain another component (peptide or pharmaceutical
agent), such that side
effects, such as, for example, reduced compatibility, occur more often.
Of the particulate contrast media, the thrombus visualization of EP 885545 is
described
but not confirmed by experiment.
In WO 02/22011, images are shown, but the latter are obtained according to T2*-

weighted flash sequences, such that the thrombi are only signal-free after the
administration of
contrast medium.
The examination of the blood of the patient for the presence of an elevated
concentration
of D-dimers has recently gained considerable clinical importance. According to
associated
studies, a concentration of less than 500 ~g/1 of D-dimers in the blood rules
out the presence of a
thromboembolism with very high probability (Wells, P. S. Brill-Edwards, P.,
Stevens, P. et al. A
Novel and Rapid Whole Blood Assay for D-Dimer in Patients with Clinical
Suspected Deep
Vein Thrombosis. Circulation 1995; 91: 2184-2187). The specificity of the D-
dimer detection
is low, however, such that a thrombosis cannot be deduced from an increase of
the concentration
in the blood.
Obtaining images with the aid of nuclear magnetic resonance (MRI) is a modern,
non-
invasive radiological process, which makes possible the visualization of
physiological and
pathophysiological structures with a very good space and time resolution. In
the diagnosis of
deep femoral and pelvic vein thrombosis, the MR venography (MRV) has been
methodically
established for quite a long time as an alternative to phlebography and color-
coded Doppler



CA 02491933 2005-O1-05
sonography (FKDS) in the area of the suprapopliteal veins. In recent years,
studies on the MRV
of the deep femoral veins were also published.
From the data record of a contrast-enhanced 3D-MR-angriography, the venous
system
basically can be selectively visualized by perfusion phase subtraction and
directly visualized via
an instep vein after injection of a dilute paramagnetic contrast medium. In
the case of MR
angiography with conventional, extracellular, paramagnetic substances, a
homogeneous vascular
contrast is not always achieved, which impedes an evaluation in individual
cases. With the
expected use of highly concentrated contrast media or so-called "blood pool
agents," however,
the faster, contrast medium-supported 3D-MRA could be advantageous. With T2-
turbo-spinecho
(TSE) and Time-of Flight (TOF)-sequences (without contrast medium), an
adequate signal level
can also be achieved in the open lower field tomograph (Konig, C. et al., MR-
Venographie am
offenen Niederfeldtomographen unter Verwendung manueller Flussaugmentation [MR
Venography in the Open Lower-Field Tomograph with Use of Manual Flow
Augmentation];
Rofo, Fortschr. Geb. Rontgenstr. Neuen Bildgeb. Verfahr. 2001; 173: 810-814).
Flow-sensitive
MRA techniques, however, are less suitable for the diagnosis of thromboses,
since in veins,
especially in those with a thrombosis, the flow rate in the non-thrombosed
portion is often too
low.
The use of specific contrast media with selective concentration in certain
tissues and
organs could increase the diagnostic value of the MR imaging considerably.
Contrast medium
preparations with selective concentration in intravascular thrombi were able
to detect location
and degree of the disease at an early time and thus to make possible a
targeted therapy and
prophylaxis.
In addition, there is therefore a need for a compatible, powerful contrast
medium for
visualizing arterial and venous thromboses.
The object of this invention was therefore to make available contrast media
for the
visualization of intravascular thrombi in MR imaging that meet the
requirements for
Selective concentration



CA 02491933 2005-O1-05
6
High compatibility
Strong enhancement
Complete elimination
and good water solubility.
It has now been found, surprisingly enough, that perfluoroalkyl-containing
metal
complexes, which have a critical micelle formation concentration < 103 mol/1,
a hydrodynamic
micelle diameter (2 Rh > 1 nm) and a proton relaxivity in plasma (R~) >
101/mmol~s, are very
well suited as contrast media in MR imaging for visualizing intravascular
thrombi.
Compounds with these properties were already described in WO 02/13874 as
diagnostic
agents for plaque imaging with the aid of MR technology.
The MR images clearly show, however, that plaques and thrombi can be
distinguished
clearly from one another. This is therefore important, since thrombi in the
young stage can be
mobile and can result in lethal embolisms.
For the following tests, the gadolinium complexes were used, since the
gadolinium of all
paramagnetic ions has the greatest influence on the signal amplification in
the MRI.
In an in-vitro test (binding to a fibrin gel), it was possible to prove that
the compounds
according to the invention bind to the fibrin gel at a concentration of 0.01
mmol of Gd/I to 79%
and at a concentration of 0.1 mmol of Gd/I to 39% and thus make possible a
reliable
differentiation from plaque.
In addition, the contrast behavior of the compounds according to the invention
was also
examined in vivo. In rabbits with photochemically induced thrombus (PIT; i.v.
injection of rose-
Bengal and irradiation with xenon light), a considerable enhancement in the
induced thrombus
could be observed at various times after intravenous administration of 0.1
mmol of Gd/kg of
body weight of the compound according to the invention (2 to 48 hours p.i.)
with T1-weighted
sequences. At the time of 24 hours p.i., the gadolinium concentration in the
thrombus was about
4 x higher compared to the blood.
For the MRI imaging, gadolinium concentrations of at least 50 p.mo/1 and at
most 2500



CA 02491933 2005-O1-05
7
~mol/1 are required in the thrombus, where the concentration of the compound
is carried out.
The imaging can be done after 15 minutes or up to 48 hours after injection of
the compounds
according to the invention. Since primarily the TI-relaxation times of the
tissue are influenced
with the gadolinium complexes according to the invention, TI-weighted
sequences are best able
to detect an enhancement in the thrombus.
Amphiphilic compounds that as a nonpolar portion have a perfluoroalkyl side
chain in the
molecule that is optionally connected via a lipophilic linker with the total
molecule are defined as
perfluoroalkyl-containing metal complexes that are suitable for use according
to the invention.
The polar portion of the compounds according to the invention is fonmed by one
or more metal
complexes and optionally present additional polar groups.
In aqueous systems, these amphiphilic molecules show the properties that are
characteristic of standard surfactants (such as, e.g., sodium dodecylsulfate,
SDS). They thus
reduce the surface tension of water. By tensiometry, the so-called CMC
(critical micelle
formation concentration in mol/1) can be determined. In this respect, the
surface tension is
determined based on the concentration of the substance to be measured. The CMC
can be
calculated from the plot of the surface tension function (c) that is obtained.
The critical micelle
formation concentration of the compounds according to the invention must be <
I 0-3 mol/1,
preferably < 10~ mol/1.
The amphiphilic compounds according to the invention are associated in
solution and are
present as aggregates. The size (2 Rh) of such aggregates (e.g., micelles,
rods, wafers, etc.) can
be determined with the aid of photon-correction spectroscopy (PCS).
As a second criterion, the hydrodynamic micelle diameter 2 Rh, which must be >
1 nm, is
therefore used. In particular, those perfluoroalkyl-containing metal complexes
according to the
invention whose 2 RH >_ 3 nm, quite especially preferably > 4 nm, are
suitable.
Both the determination of the CMC and the photon correlation spectroscopy are
described
in H.-D. Dorfler, "Grenzflachen- and Kolloidchemie [Interface and Colloid
Chemistry],"
Weinheim, New York, Basel, Cambridge, Tokyo, VSH 1994.



CA 02491933 2005-O1-05
8
As a third criterion, the proton-relaxivity in plasma (R') at 40°C and
a field strength of
0.47 tesla are used. The relaxivity, which is indicated in [l/mmol~s], is the
quantitative
measurement for the shortening of relaxation time T' of the protons. For the
purpose according
to the invention, the relaxivity must be as high as possible and > 10
I/mmol~s, preferably > 13
1/mmol~s, especially preferably > 151/mmol~s.
Relaxivity R' [l/mmol~s] of the MR-contrast media according to the invention
was
determined with the Minispec P 20 device of the Bruker Company. The
measurements were
taken at 40°C and a field strength of 0.47 tesla. Eight measuring
points were recorded by each
T 1-sequence: 180°-TI-90°, inversion recovery. As a medium,
bovine plasma of the Kraeber
Company was used. The contrast medium concentrations [mmol/I] in the batches
were between
0.30 and 1.16.
In an embodiment of this invention, the compounds of general formula I
according to
claims 8 to 11 are used as preferred compounds. In this case, these are known
compounds that
are described in WO 97/267017. Their production can also be found in this WO
publication.
Surprisingly enough, it has been shown that these compounds are also very well
suited as MRI-
contrast media for visualization of thrombi. As quite especially preferred
compounds, metal
complexes MK 2, 3 and 4, as well as MK 8, 9, 10 and 11 (cf. also Table 1 ) are
used.
In another embodiment of this invention, those compounds of general formula Ia
according to claims 12 to 21 are used as preferred compounds. These compounds
are known and
are described in WO 99/01161. Their use as MRI contrast media for
visualization of thrombi
still had not been described to date. Of these compounds, quite especially
preferably metal
complex MK 12 (cf. Table 1) is used.
In another preferred embodiment of the invention, the macrocyclic
perfluroalkyl
compounds of general formula Ib
~,,
K-~---L'-RR (Ib)
in which



CA 02491933 2005-O1-05
9
K means a complexing agent or a metal complex of general formula IIb
(Ilb)
whereby
R' stands for a hydrogen atom or a metal ion equivalent of atomic numbers
23-29, 42-46 or 58-70,
R2 and R3 stand for a hydrogen atom, a C,-C~ alkyl group, a benzyl group, a
phenyl group, -CH20H or -CHZ-OCH3, and
UZ stands for radical L', whereby L' and U2, independently of one another,
can be the same or different,
A' means a hydrogen atom, a straight-chain or branched C,-C3o alkyl group,
which
optionally is interrupted by 1-1 S oxygen atoms, and/or optionally is
substituted
with 1-10 hydroxy groups, 1-2 COOH groups; a phenyl group, a benzyl group
and/or 1-5 -OR9 groups, with R9 in the meaning of a hydrogen atom or a C,-C~
alkyl radical, or -L'-RF,
L' means a straight-chain or branched C,-C3o-alkylene group, which optionally
is
interrupted by 1-10 oxygen atoms, 1-S -NH-CO groups, 1-5 -CO-NH groups, by a
phenylene group optionally substituted by a COOH group, 1-3 sulfur atoms, 1-2
-N(B')-S02 groups and/or 1-2 -S02-N(B') groups with B' in the meaning of A',
an NHCO group, a CONH group, an N(B')-SOZ group or an -SOZ-N(B')
group andlor optionally is substituted with radical RF, and
RF means a straight-chain or branched perfluorinated alkyl radical of formula
C"FZnE,
whereby n stands for numbers 4-30, and



CA 02491933 2005-O1-05
E stands for a terminal fluorine atom, chlorine atom, bromine atom, iodine
atom or a hydrogen atom,
and optionally present acid groups optionally can be present as salts of
organic and/or inorganic
bases or amino acids or amino acid amides, as they and their production are
disclosed and
defined in WO 02/13874,
can be used.
According to the invention, metal complexes MK 17, MK I 8, MK 19, MK 21, and
MK
23 (cf. Table 1 ) are quite especially preferably used.
These compounds of general formula Ib are very well suited as MRI contrast
media for
visualizing thrombi.
In another preferred embodiment of the invention, the perfluoroalkyl-
containing
complexes with sugar radicals of general formula Ic (see WO 02/13874)
(K~~_G _ ~Z_RF~n~
(Ic)
~~R~,
in which
R represents a mono- or oligosaccharide radical bonded by the 1-OH- or 1-SH-
position,
RF is a perfluorinated, straight-chain or branched carbon chain with the
formula
-C"F2"E, in which E represents a terminal fluorine; chlorine, bromine, iodine
or
hydrogen atom, and n stands for numbers 4-30,
K stands for a metal complex of general formula IIc,



CA 02491933 2005-O1-05
ll
COOR'
~,r
,/~'h! Ra f~ 0
f ~ ~1~U
9
R OOC
~, 0
COOK'
(Ila)
in which
R~ means a hydrogen atom or a metal ion equivalent of atomic numbers 23-29, 42-
46
or 58-70,
provided that at least two R' stand for metal ion equivalents,
R2 and R3, independently of one another, represent hydrogen, C,-C~ alkyl,
benzyl, phenyl,
-CH20H or -CH20CH3, and
U represents -C6H4-O-CH2-c~, -(CHz)~_5-w, a phenylene group, -CH2-NHCO-CHz-
CH(CHZCOOH)-C6H4-w, -C6H4-(OCHZCHz)o_1-N(CH2COOH)-CHZ-c~, or a C,-
C,2 alkylene group or C~-C,2-C6H4-O group optionally interrupted by one or
more
oxygen atoms, 1 to 3 -NHCO groups or 1 to 3 -CONH groups and/or substituted
with 1 to 3 -(CH2)o_5 COOH groups, whereby c~ stands for the binding site
to -CO-,
or
of general formula IIIc
DOOR'
4
COOK
R'OOC''r / l!
~,N
COO R'
tilic)



CA 02491933 2005-O1-05
12
in which R' has the above-mentioned meaning, R'° represents hydrogen or
a metal ion equivalent
mentioned under R~, and U' represents -C6H4-O-CH2-ca, whereby w means the
binding
site to -CO-,
or of general formula IVc
COOR'
~N R
R'OOC'N
N O
GoOR'
~iVc)
in which R' and R2 have the above-mentioned meaning
or of general formula VcA or VcB
N,.--~-Ca-~
''-Cad'
R'ObC'~N
N~Ct)aR~
eaaR
c~~>
Nr-caoR'
o ' ~-cooR~
,,~.-G~.,-- jN
~~~cooR~
COOK
N~)
in which R~ has the above-mentioned meaning,



CA 02491933 2005-O1-05
13
or of general formula VIc
R't7C7C---,~, ,r'--~ ,..-wC0-.,~.
R'000-~'~N N'~-~.~Ot7R'
(Vlc)
in which R' has the above-mentioned meaning,
or of general formula VIII
R'OOC--~, O
R'OC?C---''°~ U'
R'OOC--~,.,N
R'OOC--~
(1/f l~)
in which R' has the above-mentioned meaning, and
U~ represents -C6H4-O-CHZ-w, whereby w means the binding site to -CO-
or of general formula VIIIc
~00!R'
'N
R'OOC''
N
s
~Ot7R ~ilillc)
in which R' has the above-mentioned meaning,
and in radical K, optionally present free acid groups optionally can be
present as salts of
organic and/or inorganic bases or amino acids or amino acid amides,



CA 02491933 2005-O1-05
14
G for the case that K means metal complexes IIc to VIII, represents a radical
that is
functionalized in at least three places and is selected from the following
radicals
a) to j)
(a1
a ~~-~. (CHZ),~ ~---GO --~.,~ Y
NH
(a2~
H
a-~ H (CH~)d-C-C
NH
1



CA 02491933 2005-O1-05
15
Y ~-~O-~-f~Hz~~ ~ ~- ~
NH
a
fe)
ac
~' N
v~,.- --,-"w,. Y
~N~
a
f~)



CA 02491933 2005-O1-05
16
~~)
',.,
~.,~.--~ ~~...,. «
~1~-i-Ct~-~---~Gl-~~~ ~~«
N-~'Y
w
-~U-C-~CH~a~ ~'~'w-oc
filw



CA 02491933 2005-O1-05
17
NH
NH--CC-GH-(CH2); NH-~''~
N-CC~--~H--(CH2~~ NHw~~~''a
NH
NH--CQ-~H-~GH2); NH-~~~'
NH
a~.M..--~.-.iCH~,~~-C-~---CO.C,...,-(CH~~~-~--.-"~
(~H
Y
cx
f~
(~.,-~-_-~CH~~~a C-C ~ N~-~wY
hi ~1H
a



CA 02491933 2005-O1-05
18
ac,~lv~-I_~CH -00-~nAY
and
G for the case that K means metal complex VIIIc, represents a radical that is
functionalized in at least three places and is selected from k) or 1),
-NH---(~H~~;~ CH-GO-~''ac
i~H
Y
a~-Cp~--CHI ~H~-~H~ CO-~'Y
f~H
whereby a means the binding site of G to complex K, ~ is the binding site of G
to radical
Y, and y represents the binding site of G to radical Z,
Y means -CH2, 8-(CH2),_SCO-Vii, ~i-(CH2),_SCO-8, 8-CHz-CHOH-CO-(3 or 8-
CH(CHOH-CHZOH)-CHOH-CHOH-CO-(3, whereby 8 represents the binding site
to sugar radical R, and ~ is the binding site to radical G,
Z stands for



CA 02491933 2005-O1-05
19
~ ~~ N-S0~
y-CaCH~-N(C~H3}-S(3~-s,
y~CC3CH,~-C7-(CH~O~-s,
a a
~ON.~~.N--S02 ~
or
y - NHCHzCH2-O-CHZCHz-~
whereby y represents the binding site of Z to radical G, and ~ means the
binding
site of Z to perfluorinated radical RF
and
1', m', independently of one another, mean integer I or 2, and
p' means integers 1 to 4,
can be used.
As quite especially preferred compounds of general formula Ic, metal complex
MK I 3 of
Table I according to the invention is used.
In another preferred embodiment of the invention, the perfluoroalkyl-
containing
complexes with polar radicals of general formula Id (see WO 02/13874) are used
(~~~~C~ RFlm~ (Id)
t ~~~z
in which
RF is a perfluorinated, straight-chain or branched carbon chain with formula -
C"FZ~E,
in which E represents a terminal fluorine, chlorine, bromine, iodine or
hydrogen
atom, and n stands for numbers 4-30,



CA 02491933 2005-O1-05
K stands for a metal complex of general formula IId,
COON'
Ra O
i
~'~-u~
(Il~d)
in which
R' means a hydrogen atom or a metal ion equivalent of atomic numbers 23-29, 42-
46
or 58-70,
provided that at least two R' stand for metal ion equivalents,
RZ and R3, independently of one another, represent hydrogen, C,-C~ alkyl,
benzyl, phenyl,
-CH20H or -CHZOCH3, and
U represents -C6H4-O-CH2-w-, -(CHz),_5-c~, a phenylene group, -CH2-NHCO-CH2-
CH(CHZCOOH)-C6H4-w-, -C6H4-(OCH2CH2)o_~-N(CHzCOOH)-CHZ-w, or a C,-
C,2 alkylene group or C~-C~2-C6H4-O group optionally interrupted by one or
more
oxygen atoms, 1 to 3 -NHCO groups, 1 to 3 -CONH groups and/or substituted
with 1 to 3 -(CH2)o-sCOOH groups, whereby w stands for the binding site to -CO-
,
or



CA 02491933 2005-O1-05
21
of general formula IIId
C00 R'
N ~ GOOR I
.,...~.,. ,.._-
R'OOC'~~ ~ U
N
G~70R'
(Ilid)
in which R1 has the above-mentioned meaning, R4 represents hydrogen or a metal
ion equivalent
mentioned under R', and U' represents -C6H4-O-CH2-w-, whereby c~ means the
binding site to
-CO-,
or of general formula IVd
GOOK'
R'QOC''
~GC?4!;'
(llfd~
in which R' and Rz have the above-mentioned meaning,



CA 02491933 2005-O1-05
22
or of general formula VdA or VdB
~N~C 0
R'OOC"'~,
~N~C0~0R~
G04R
~'ciA)
Nr-Cta0l~'
p ~ '~-COOK'
~-C~
N.r-CO UR'
'~-CO OR'
(~dB3
in which R' has the above-mentioned meaning,
or of general formula VId
R'OOC--~,.' f"~ ~,,---CO-..~,,~,
R'OC~C--~''N ~'~,-.-C(7E.)R'
(IIId~
in which R' has the above-mentioned meaning,
or of general formula VIId
R'O CSC-,-,~ ~0
R'UC7C-''N '~
R'C~OC-,~N
R'OOC-~'
(Vlid)



CA 02491933 2005-O1-05
23
in which R~ has the above-mentioned meaning, and
U ~ represents -C6H4-O-CHZ-c~-, whereby ca means the binding site to -CO-,
and in radical K, optionally present free acid groups optionally can be
present as salts of
organic and/or inorganic bases or amino acids or amino acid amides,
G represents a radical that is functionalized in at least three places and is
selected
from the following radicals a) to i)
t~~ ?
NH
(a2)
F
a~H (~H~~j4 C-CU-rwlr ~3
~JH
I
Y
(~~
'Y ,n~..~CO-~-(~Hz)~ ~ w~- ~
~H
a



Image



CA 02491933 2005-O1-05
(fib
«,~»,...--~-~cH~~~-c-~-co-C--~cH~~~ H--~~
~o
~!H
ts)
HM N NH
~f 1 Y wQ~to~x~a-~~"',w'~ . (i) ~ ~wV Co-(Cliz)xw ~r~co.",~r
~H
a a
whereby a means the binding site of G to complex K, (3 is the binding site of
G to radical R, and
y represents the binding site of G to radical Z
Z stands for
~-hl ~ N-~~~ 8
r..G~O~Hzo~CHz~.
whereby y represents the binding site of Z to radical G, and ~ means the
binding
site of Z to perfluorinated radical RF,
R represents a polar radical that is selected from complexes K of general
formulas
IId to VIId, whereby R' here means a hydrogen atom or a metal ion equivalent
of



CA 02491933 2005-O1-05
26
atomic numbers 20, 23-29, 42-46 or 58-70,
and radicals RZ, R3, R4, U and U' have the above-indicated meaning,
or
the folic acid radical
or
means a carbon chain with 2-30 C atoms that is bonded to radical G via -CO- or
SOZ- or a direct bond, and is straight or branched, saturated or
unsaturated, optionally interrupted by 1-10 oxygen atoms, 1-5 -NHCO groups, 1-
5
-CONH groups, 1-2 sulfur atoms, 1-5 -NH groups or 1-2 phenylene groups, which
optionally can be substituted with 1-2 OH groups, 1-2 NH2 groups, 1-2
-COOH groups, or 1-2 -S03H groups,
or
optionally substituted with 1-8 OH groups, 1-5 -COOH groups, 1-2 S03H groups,
1-5 NH2 groups, or I-5 C,-C4 alkoxy groups, and
1', ml, p2, independently of one another, mean integer 1 or 2.
Especially preferred compounds of general formula Id are those with
macrocyclic
compound K of general fornmla IId, IIId, VdB or VIId.
Metal complex MK12 of Table 1 according to the invention is used as a quite
especially
preferred compound of general formula Id.
In another preferred embodiment of the invention, galenical formulations can
be used that
contain paramagnetic and diamagnetic perfluoroallkyl-containing substances.
The paramagnetic
and diamagnetic substances are preferably present in a dissolved state in an
aqueous solvent.
As paramagnetic, perfluoroalkyl-containing compounds, all above-mentioned
metal
complexes of general formulas I, Ia, Ib, Ic and/or Id according to the
invention can be used in the
formulations.
The diamagnetic perfluoroalkyl-containing substances are those of general
formula XX
(see WO 02/13874):



CA 02491933 2005-O1-05
27
RF_L2_B2 (XX)
in which RF represents a straight-chain or branched perfluoroalkyl radical
with 4 to 30 carbon
atoms, LZ stands for a linker, and BZ stands for a hydrophilic group. Linker
L2 is a direct bond,
an -SOZ group, or a straight-chain or branched carbon chain with up to 20
carbon atoms, which
can be substituted with one or more -OH, -COO, or -S03 groups and/or
optionally contains one
or more -O-, -S-, -CO-, -CONH-, -NHCO-, -CONR9-, -NR9C0-, -S02-, -P04-, -NH-
or -NR9
groups, an aryl ring or a piperazine, whereby R9 stands for a C, to CZO alkyl
radical, which in turn
can contain one or more O atoms, and/or can be substituted with -COO- or S03
groups.
Other suitable diamagnetic perfluoroalkyl-containing compounds are conjugates
that
consist of cyclodextrin and perfluoroalkyl-containing compounds. These
conjugates consist of
a-, (3- or y-cyclodextrin and compounds of general formula XXII (see WO
02/13874)
A ~ _L3_RF (XXII)
in which A' stands for an adamantane, biphenyl or anthracene molecule, L3
stands for a linker,
and RF stands for a straight-chain or branched perfluoroalkyl radical with 4
to 30 carbon atoms.
Linker L3 is a straight-chain hydrocarbon chain with 1 to 20 carbon atoms,
which can be
interrupted by one or more oxygen atoms, one or more CO-, S02-, CONH-, NHCO-,
CONR-,
NRCO-, NH- or NR groups or a piperazine, whereby R is a C,-CS alkyl radical.
In summary, it has been detemlined that as quite especially preferred
compounds,
gadolinium complexes MK I-30 that are presented in Table 1 meet the criteria
according to the
invention. The physical parameters of these metal complexes are presented in
Table I .
Both the paramagnetic compounds of general formulas I, Ia, Ib, Ic and Id
according to the
invention and the formulations that consist of paramagnetic and diamagnetic
perfluoroalkyl-
containing substances according to the invention are extremely well suited as
contrast media in
MR-imaging for visualization of thrombi.



CA 02491933 2005-O1-05
28
Table 1:
Metal Complexes (MIA that are Quite Especially Preferably Used According to
the
Invention, Their Origin and Their Physicochemical Parameters
Com lex Ori in Exam le R' [l : CMC[mol/1] 2 Rh [nm]
No. mmol's]


MK 1 WO 99/0116118 23.0 1.5 10 3.5


MK 2 WO 97/260171 29.7 1.0 10-5 31.5


MK 3 WO 97/260172 33.0 2.3 10-5 14.0


MK 4 WO 97/260173 27.5 1.44 10-5 3.2


MK S WO 99/0116125 15.1 3.1 10- 7.0


MK 6 WO 97/2601731 26.0 9.8 10 4.3


MK 7 WO 99/0116112 21.4 1.81 10 4.2


MK 8 WO 97/2601733 35.7 1.86 ' 10- 4.6


MK 9 WO 97/2610735 34.0 3.25 ~ 10- 4.3


MK 10 WO 97/2601734 24.9 7.06 ' 10- 3.2


MK 11 WO 97/2601732 24.8 2.88 ' 10- 35.5


MK 12 WO 99/011611 19.5 8.9 ' 10 2.2


MK 13 WO 02/1387421 15.9 2.5 ' 10 4.4


MK 14 WO 02/1387454 21.3 3.9 10- 4.9


MK 15 WO 99/0116114 19.3 8.7 ~ 10- 3.2


MK 16 WO 00/567237 21.0 2.8 ' 10- 4.3


MK 17 WO 02/138746 13.3 2.65 ' 10- 6.0


MK 18 WO 02/138742 19.6 3.9 - 10~ 4.4


MK 19 WO 02/138745 30.3 5.2 ' 105 3.0


MK 20 WO 00/567234 21.9 4.6 10- 5.5


MK 21 WO 02/138743 21.2 2.92 10- 2.5


MK 22 WO 00/567237 27.8 4.4 ' 10 5.7


MK 23 WO 02/138741 25.7 7.9 ' 10- 5.4


MK 24 WO 99/011611 13.9 6.3 ' 10 10.0


MK 25 WO 99/011615 21.3 1.4 10 3.5


MK 26 WO 02/1387457 22.8 4.3 ' 10- 5.2


MK 27 WO 97/2501738 30.5 1.07 ' 10- 7.4


MK 28 This 1 27.9 8.1 ' 10~ 4.7
a lication


MK 29 This 2 17.7 7.6 105 4.8
a lication


MK 30 This 3 27.9 7.0 ~ 10- 7.9
a lication





CA 02491933 2005-O1-05
29
CMC: Critical Micelle Formation Concentration
2 Rh: Hydrodynamic Micelle Diameter
R' : Relaxivity
The measurements were carried out in plasma at 40°C and a field
strength of 0.47 Tesla.
Example 1
a) 6-Benzyloxycarbonyl-2-N-2H,2H,4H,4H,SH,SH-3-oxa-perfluorotridecanoyl-L-
lysine
methyl ester
2 drops of dimethylformamide are added to the solution of SO g (95.8 mmol) of
2H,2H,4H,4H,SH,SH-3-oxaperfluorotridecanoic acid (produced from 2H,2H,3H,3H-
perfluorodecanol and bromoacetic acid-t-butyl ester with subsequent ester
cleavage) in 250 ml of
thionyl chloride, and it is refluxed for 5 hours. Then, it is concentrated by
evaporation in a
vacuum, the residue is taken up in 250 ml of dichloromethane, and the solution
is added in drops
at 0°C while being stirred to the solution of 34.74 g ( 105.0 mmol) of
6-N-benzyloxycarbonyl-L-
lysine-methyl ester, hydrochloride (commercially available products; Bachem)
as well as 46.85
ml (350 mmol) of triethylamine in 400 ml of dichloromethane. It is allowed to
stir overnight,
mixed with 1 liter of 2N hydrochloric acid, the organic phase is shaken out,
the water phase is
extracted twice with 100 ml of dichloromethane each, the solution is dried on
sodium sulfate,
dessicant is filtered out, and it is concentrated by evaporation in a vacuum.
The crude product is
purified on silica gel by chromatography. As an eluant, a mixture that
consists of
dichloromethane with the addition of 3% ethanol is used.



CA 02491933 2005-O1-05
'The product is obtained as a colorless gel after the concentration by
evaporation.
Yield: 67.0 g (87.6% of theory)
Elementary analysis:
Cld.: 40.61 C 3.41 H 40.45 F 3.51 N
Fnd.: 40.48 C 3.54 H 40.61 F 3.37 N.
b) 2-N-2H,2H,4H,4H,SH,S,H-3-Oxaperfluorotridecanoyl-L-lysine methyl ester,
hydrochloride
I 0 g of catalyst (Pd I 0%/C) is added to a solution of 63.5 g (79.5 mmol) of
the title
substance of Example 1 a) in a mixture that consists of 500 ml of methanol and
90 ml of 1 N
hydrochloric acid, and it is hydrogenated until one equivalent of hydrogen is
taken up at normal
pressure and room temperature. Catalyst is filtered out, the latter is washed
3 times with 50 ml
each of hot methanol, and the combined solutions are concentrated by
evaporation. The residue
is dissolved in methanol and brought to crystallization by adding diisopropyl
ether.
The title compound is obtained in colorless crystals.
Yield: 55.70 g (quantitative)
Elementary analysis:
Cld.: 32.56 C 3.16 H 5.06 CI 46.09 F 4.00 N
Fnd.: 32.44 C 3.28 H 4.95 Cl 46.21 F 4.1 I N



CA 02491933 2005-O1-05
3I
c) 6-N-3,6,9.12.15-Pentaoxa-hexadecanoyl-2-N-2H,2H,4H,4H,SH,SH-3-oxa-
perfluorotridecanoyl-I-lysine-methyl ester
13.3 I g (50.0 mmol) of 3,6,9,12,15-pentaoxahexadecanoic acid (commercially
available
product) is dissolved in 100 ml of thionyl chloride, mixed with two drops of
dimethylformamide
and stirred overnight at room temperature. Then, it is heated for one hour to
65°C, excess thionyl
chloride is removed in a rotary evaporator, and the residue is taken up in I
50 ml of
dichloromethane. This solution is added in drops at 0°C to the solution
of 35.04 g (50.0 mmol)
of the title compound of Example Ib) and I5. I 8 g (150 mmol) of triethylamine
in 350 ml of
dichloromethane. Then, it is allowed to stir for 72 hours at room temperature.
It is concentrated
by evaporation, and the product is obtained by column chromatography on silica
gel. As an
eluant, a mixture that consists of dichloromethane/ethanol 9: I is used. The
title compound is
obtained as a viscous, light yellow oil.
Yield: 37.0 g (81.1 % of theory)
Elementary analysis:
Cld.: 39.48 C 4.53 H 35.39 F 3.07 N
Fnd.: 39.61 C 4.50 H 35.50 F 3.16 N
d) 6-N-3,6,9,12,1 S-Pentaoxahexadecanoyl-2-N-2H,2H,4H,4H,SH,SH-3-oxa-
perfluorotridecanoyl-L-lysine
17.90 g of the title compound of Example 1 c) is stirred overnight in a
mixture that



CA 02491933 2005-O1-05
32
consists of 50 ml of methanol and 25 ml of 2N sodium hydroxide solution. It is
acidified with
2N hydrochloric acid, concentrated by evaporation in a vacuum, and the residue
is extracted 5
times with 50 ml each of tetrahydrofuran/ethyl acetate 2:1. The combined
extracts are dried on
sodium sulfate. Desiccant is filtered out, and the solution is concentrated by
evaporation. The
residue is purified by column chromatography on silica gel. As a mobile
solvent, a mixture that
consists of dichloromethane/methanol and water in a ratio of 160:40:1 is used.
The title
compound is obtained as a waxy, light yellow-colored residue.
Yield: 14.7 g (83.4% of theory)
Elementary analysis:
Cld.: 38.76 C 4.37 H 35.94 F 3.I2 N
Fnd.: 38.87 C 4.25 H 36.07 F 3.21 N
e) 6-N-3,6,9,12,15-Pentaoxahexadecanoyl-2-N-2H,2H,4H,4H,5H,SH-3-
oxaperfluorotridecanoyl-L-lysine-N- { 1,4, 7-tris[carboxylatomethyl]-1,4, 7,10-

tetraazacyclododecane-10-(2-hydroxy-3-yl), gadolinium complex}-amide
8.0 g (8.9 mmol) of the acid that is produced under 1 d) as well as 2.05 g (
17.8 mmol) of
hydroxysuccinimide are dissolved in 50 ml of dimethylformamide and mixed at
0°C with 4.60 g
(22.25 mmol) of dicyclohexylcarbodiimide. It is stirred for 10 more minutes at
0°C and then for
another 2 hours at room temperature. After being cooled again to 0°C, a
solution that consists of
3.93 g (6.65 mmol) of the gadolinium complex of 10-(3-amino-2-hydroxy-propyl)-
1,4,7-tris-
(carboxymethyl)-1,4,7,10-tetraazacyclododecane (WO 9S/17451) as well as O.S8 g
(13.7 mmol)
of lithium chloride and 2.77 g (27.4 mmol) of triethylamine in 40 ml of
dimethyl sulfoxide is



CA 02491933 2005-O1-05
33
added. It is allowed to stir for two days at room temperature, mixed with 650
ml of acetone, and
the solution is poured onto 21 of methyl-t-butyl ether. It is stirred for
about 30 more minutes,
and then solid is suctioned out. The solid is dissolved in distilled water and
treated with
activated carbon. The solution is filtered, concentrated by evaporation in a
vacuum, and the
residue is chromatographed on silica gel. As an eluant, a mixture that
consists of methanol and
dichloromethane is used in a 2:1 ratio.
Yield: 5.47 g (54.5% of theory)
Water content: 7.3%
Elementary analysis (relative to anhydrous substance):
Cld.: 37.97 C 4.71 H 10.81 Gd 22.19 F 6.74 N
Fnd.: 38.16 4.83 10.72 22.32 6.83
Example 2
a) 10-(3-Carboxy-3-yl-propionic acid)-1,4,7,10-tetraazacyclododecane
150 g (761 mmol) of bromosuccinic acid is neutralized with sodium hydroxide
solution
(10%), and the solution is evaporated to the dry state. 65.55 g (380 mmol) of
cyclene is
dissolved in 300 ml of distilled water and mixed with the bisodium salt (from
150 g of
bromosuccinic acid = 761 mmol). It is heated to SO°C and allowed to
stir overnight. The
solution is then evaporated to the dry state and co-distilled with ethanol.
The residue is taken up
in butanol and extracted with water. The aqueous phase is concentrated by
evaporation and
chromatographed on silica gel. As an eluant, mixtures that consist of methanol
with ammonia
(20:1 - 2:1 ) are used. The product-containing fractions are combined and
evaporated to the dry
state.
Yield: 54.8 g (50.4% of theory)



CA 02491933 2005-O1-05
34
Elementary analysis:
Cld.: C 50.34 H 7.74 N 19.57
Fnd.: C 50.46 H 7.83 N 19.69
b) 1,4,7-Tris(carboxymethyl)-10-(3-carboxy-3-yl-propionic acid)-1,4,7,10-
tetraazacyclododecane
11 g (38.14 mmol) of 10-(3-carboxy-3-yl-propionic acid)-1,4,7,10-
tetraazacyclododecane
is dissolved in 60 ml of distilled water and mixed with 18.03 g ( 190.74 mmol)
of chloroacetic
acid. It is then heated to 70°C, and the pH is kept between 9 and 10 by
adding sodium
hydroxide solution (32%). It is allowed to stir overnight at 70°C, then
it is set again at a pH of
10, and 7.2 g (76.19 mmol) of chloroacetic acid is added to it. It is stirred
for 3 more hours at
70°C. It is evaporated to the dry state, evaporated with methanol,
taken up in methanol, and salts
are filtered out. The filtrate is concentrated by evaporation and
chromatographed on an ion-
exchange column Amberlite 252 C with water/ammonia as an eluant. The product-
containing
fractions are combined, concentrated by evaporation, taken up again in
distilled water and freeze-
dried. The title compound is obtained as a white foam.
Yield: 13.12 g (82.3% of theory)
Water content: 9.6%
Elementary analysis (relative to anhydrous substance):
Cld.: C 46.75 H 6.54 N 12.12
Fnd.: C 46.87 H 6.62 N 12.24



CA 02491933 2005-O1-05
c) Gadolinium complex of 10-{1-carboxy-2-carbonyl-[piperazin-I-yl-4-
(perfluorooctylsul fonyl)] } -ethyl-1,4, 7-tris(carboxymethyl)-1,4,7, I 0-
tetraazacyclododecane, sodium salt
10.0 g ( 16.21 mmol) of the gadolinium complex tetraazacyclododecane (produced
from
the ligand by complexing with gadolinium oxide) and 3.0 g of lithium chloride
are dissolved
while being heated slightly in 100 ml of dimethyl sulfoxide. After cooling to
room temperature,
9.21 g ( 16.21 mmol) of perfluorooctylsulfonylpiperazine is added. Then, it is
cooled to 0°C, and
12.3 g (46.63 mmol) of EEDQ (1,2-dihydro2-ethoxyquinoline-1-carboxylic acid
ethyl ester) is
added, and it is stirred overnight at room temperature. The reaction is poured
into a mixture that
consists of 800 ml of methyl-t-butyl ether and 100 ml of acetone, and it is
stirred. The precipitate
is purified by chromatography on silica gel. A mixture that consists of
dichloromethane/methanol and ammonia in a ratio of 2:2.1 is used as an eluant.
The product-
containing fractions are combined and concentrated by evaporation. The residue
is dissolved in
200 ml of distilled water, set at a pH of 7.2 with sodium hydroxide solution
and freeze-dried.
The title compound is obtained as a white foam.
Yield: 7.64 g (39% of theory)
Water content: 7:8%
Elementary analysis (relative to anhydrous substance):
Cld.: 30.31 C 2.80 H 27.17 F 13.23 Gd 7.07 N 1.93 Na 2.70 S
Fnd.: 30.42 C 2.91 H 27.04 F 13.29 Gd 7.15 N 2.04 Na 2.59 S



CA 02491933 2005-O1-05
36
Example 3
a) 1,4,7-Tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-10-[2-hydroxy3-
(N-
benzyloxycarbonyltriglycidyl)-)-aminoJ-propyl, gadolinium complex
12.68 g (39.121 mmol) of N-benzyloxy-carbonyltriglycine (commercially
available
product, Bachem) is dissolved in 100 ml of dimethylformamide and mixed with
9.03 g (78.42
mmol) of N-hydroxysuccinimide. It is cooled to 0°C, and then 32.36 g (
156.84 mmol) of
dicyclohexylcarbodiimide is added to it. It is stirred for 20 minutes at
0°C and then for another 3
hours at room temperature. This suspension is then added to the solution,
cooled to 0°C, that
consists of 15 g (26.14 mmol) of the gadolinium complex of 10-(3-amino-2-
hydroxy-propyl)-
1,4,7-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane (produced according
to WO 95/17451)
in 40 ml of distilled water and 15 ml ( 14 mmol) of triethylamine in 60 ml of
isopropanol while
being stirred. After addition is completed, it is stirred for 3 hours at room
temperature. Then,
urea is filtered out, it is rewashed with n-butanol and concentrated by
evaporation in a vacuum.
The residue is extracted several times with water. The organic phase is dried
on sodium sulfate
and concentrated by evaporation in a vacuum. The residue is chromatographed on
silica gel. As
an eluant, a mixture that consists of dichloromethane, methanol and ammonia is
used. The
product-containing fractions are combined, concentrated by evaporation in a
vacuum, taken up
again in distilled water, and subjected to freeze-drying. The title compound
is obtained as a
white foam.
Yield: 12.94 g (56.3% of theory)
Elementary analysis:
Cld.: 42.36 C 5.16 H 17.89 Gd 12.75 N
Fnd.: 42.44 C 5.22 H 17.78 Gd 12.80 N



CA 02491933 2005-O1-05
37
b) 1,4,7-Tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-10-(2-hydroxy-
3-amino-
triglycidyl)-propyl, gadolinium complex
In a mixture that consists of 100 ml of ethanol and 30 ml of distilled water,
8.53 g (9.7
mmol) of the title compound of Example 3a) is dissolved and mixed with 2 g of
catalyst
(palladium 10% on activated carbon) as well as 3 ml of acetic acid. It is
hydrogenated until one
equivalent of hydrogen is taken up. Then, catalyst is suctioned out, it is
rewashed with ethanol,
and the solution is evaporated to the dry state in a vacuum.
The title compound is obtained as a foam.
Yield: 7.22 g (quantitative)
Elementary analysis:
Cld.: 37.08 C 5.28 H 21.11 Gd 15.04 N
Fnd.: 37.21 C 5.33 H 21.25 Gd 1 S.15 N
c) 1,4,7-Tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-10-{2-hydroxy-
3-N-
[triglycidyl-N-(2H,2H,4H,4H,SH,SH-3-oxa-perfluorotridecanoyl)]amino}-propyl,
gadolinium complex
7.60 g ( 14.55 mmol) of 2H,2H,4H,4H,SH,SH-3-oxa-perfluorotridecanoic acid is
dissolved in 90 ml of dimethylformamide and mixed with 3.35 g (29.1 mmol) of N-

hydroxysuccinimide. It is cooled to 0°C and then mixed with 11.71 g
(56.71 mmol) of
dicyclohexylcarbodiimide. A$er 20 minutes, the cooling is removed, and it is
stirred for another
3 hours at room temperature. Then, the suspension that was produced is added
while being
stirred to the solution, cooled to 0°C, of 7.22 g (9.7 mmol) of the
title compound of Example 3b)
in a mixture that consists of 5 ml (36.7 mmol) of triethylamine, 20 ml of
distilled water, and 30
ml of 2-propanol. It is allowed to stir overnight at room temperature, then
dicyclohexylurea is



CA 02491933 2005-O1-05
38
filtered out, it is rewashed with 2-propanol/distilled water 3:2 and the
combined solutions are
concentrated by evaporation in a vacuum. The residue is dissolved in a mixture
that consists of
water and butanol and extracted with butanol. The combined organic solutions
are dried and
concentrated by evaporation in a vacuum. The residue is purified by
chromatography on silica
gel. A mixture that consists of ethanol/2-propanol/concentrated ammonia in a
ratio of 15:10:1 is
used as an eluant. The product-containing fractions are combined, evaporated
to the dry state in
a vacuum, dissolved again in distilled water and freeze-dried. The title
compound is obtained as
a white foam.
Yield: 7.52 g (62.1 % of theory)
Elementary analysis:
Cld.: 33.66 C 3.55 H 25.86 F 12.59 Gd 8.97 N
Fnd.: 33.55 C 3.67 H 25.99 F 12.43 Gd 9.09 N
Example 4: Binding to a Fibrin Gel
A coagulate (fibrin gel) forms after a 30-minute incubation time (room
temperature) by
mixing fibrinogen with thrombin. The latter is mixed with 0.5 ml of PBS and
with 0.5 ml of a
solution of the title compound of Example 21, WO 02/13874, Mk 13, (0.01 and
0.1 mmol of
Gd/1), and it is incubated over 16 hours at room temperature. After the
supernatant is removed in
the fibrin gel, the unbonded portion of the compound according to the
invention is separated
from fibrin by ultrafiltration (1,200 g for 30 minutes). The gadolinium
content in the fibrin gel is
determined by means of inductively coupled plasma-atom emissions spectroscopy
(ICP-AES).
The binding of the compound according to the invention to the fibrin gel was
79.1 % for
the 0.01 mmol of Gd/1 solution and 38.5% for the 0.1 mmol of Gd/1 solution.



CA 02491933 2005-O1-05
39
Example 5: MRT Visualization (In-Vivo) of a Venous Thrombus after Intravenous
Administration of the Contrast Medium in Rabbits
'The MR imaging was carried out in rabbits with photochemically induced
thrombus
(PIT). By irradiation with xenon light (540 nm, 1,100 klux, 25 minutes) after
i.v. injection of
rose-Bengal (20 mg/kg), the thrombus formation was induced in the left femoral
vein. The blood
flow in the femoral vein was controlled by means of an ultrasound probe. The
imaging was
carned out with a Magnetom Harmony (Siemens, 1T) before (baseline) as well as
25, 40 minutes,
l, 2, 3, 4, 24, and 48 hours after intravenous administration (about 1 hour
after the thrombus
induction) of 0.1 mmol of Gd/kg of the title compound of Example 21, WO
02/13874, with use
of a phase-contrast sequence (TR/TE = 104/14 ms) as well as T1-weighted
gradient echo
sequences (MPRage: TR/TE/TI/a = 11/4/120ms/8°; and 3D flash: TR/TE/a =
S/2ms/SO°).
After the imaging, the left femoral vein (with the thrombus) was prepared
outside, fixed
in formalin, and stained for histological evaluation with hematoxylin/eosin
(HE) or
phosphotungstic acid/hematoxylin (PTAH).
In the MR imaging (MRA), the thrombus was already detectable early on (25
minutes
p.i.). Images 1 and 2 that are indicated in Figure 1 show MR images of the
pelvic region 24
hours after intravenous administration of 0.1 mmol of Gd/kg of body weight of
the compound
according to the invention in the PIT rabbit (photochemically induced
thrombus). The Tl-
weighted 3D-flash sequence illustrates a strong signal increase in the
thrombus in the area of the
left femoral vein. The blood flow in the left femoral vein is considerably
reduced (see MRI with
phase-contrast sequence).
With both staining techniques (HE and PTAH (Fig. 2, Images 3 and 4)), the red
blood
clots (thrombi) could be detected in the area of the left femoral vein. The
thrombi fill almost the



CA 02491933 2005-O1-05
entire lumen of the blood vessel. The exfoliation of the vascular endothelial
cells and the
adhesion of the thrombi is clearly visible. The intima- and adventitia nuclei
have disappeared
almost completely.
With this test, the suitability of the compounds according to the invention
could be shown
as markers for venous thrombi.
Example 6 MRT Visualization (Ex-Vivo) of a Venous Thrombus According to
Intravenous Administration of the Contrast Medium in Rabbits
The MR imaging was carried out in rabbits with photochemically induced
thrombus
(PIT). By irradiation with xenon light (540 nm, 1,100 klux, 25 minutes) after
i.v. injection of
rose-Bengal (20 mg/kg), the thrombus formation was induced in the left femoral
vein. 24 hours
after intravenous administration (about 1 hour a8er the thrombus induction) of
0.1 mmol of
Gd/kg of the title compound of Example 21 WO 02/13874, the animal was
sacrificed, and the
left femoral vein (with the thrombus) was prepared outside. The imaging of the
damaged venous
segment was carned out with a Magnetom I-Iarmony (Siemens; 1T) with use of a
T1-weighted
spin echo sequence (Tlt/TE/a = 300/12 ms/90°, with and without fat
suppression).
The induced thrombus is clearly visible in the preparation by the color
change. In
addition, blood clots are also found outside of the vessel. In the ex-vivo MR
imaging, a
considerable enhancement of the thrombi can be observed with the T1-weighted
spin echo
sequence (see Fig. 3, Images S to 7).



CA 02491933 2005-O1-05
41
Example 7: Determination of the Gadolinium Concentration in the Thrombus After
Intravenous Administration of the Contrast Medium in Rabbits
The determination of content was carried out in rabbits with photochemically
induced
thrombus (PIT). By irradiation with xenon light (540 nm, 1,100 klux, 25
minutes) after i.v.
injection of rose-Bengal (20 mg/kg), the thrombus formation was induced in the
left femoral
vein. 24 hours after intravenous administration (about 1 hour after the
thrombus induction) of
0.1 mmol of Gd/kg of the title compound of Example 21 WO 02/13874, the animal
was
sacrificed, and various organs and tissues were removed to determine the Gd
content: blood,
femoral veins (with and without thrombus), muscle. After the tissue samples
decomposed, the
gadolinium concentration (ppm) was measured by means of ICP-AES.
In the left femoral vein (with thrombus), the Gd concentration was 63 ppm;
however, it
was only 35 ppm in the control vessel. The blood clot outside of the vessel
had a high Gd
content of 166 ppm. In the blood at the time of 24 hours p.i., only a Gd
concentration of 15 ppm
was detectable, and in the non-signal-enhanced muscle, a Gd concentration of
10 ppm was
detectable.